MINT MINT, Dpt of Biology, University of Rome Tor Vergata MINT http://mint.bio.uniroma2.it/mint url>http://mint.bio.uniroma2.it/mint/ https://mint.bio.uniroma2.it MINT-[0-9]+ https://mint.bio.uniroma2.it/index.php/results-interactions/?id=${ac} The Molecular INTeraction database (MINT) is a relational database designed to store interactions between biological molecules. http://mint.bio.uniroma2.it An N-terminal acidic region of Sgs1 interacts with Rpa70 and recruits Rad53 kinase to stalled forks. An N-terminal acidic region of Sgs1 interacts with Rpa70 and recruits Rad53 kinase to stalled forks. EMBO J. (0261-4189) 2012 imex curation Hegnauer AM., Hustedt N., Shimada K., Pike BL., Vogel M., Amsler P., Rubin SM., van Leeuwen F., Guénolé A., van Attikum H., Thomä NH., Gasser SM. Only protein-protein interactions in vitro In vitro itc isothermal titration calorimetry ITC ITC itc experimental particp experimental participant identification experimental particp Hegnauer AM., Hustedt N., Shimada K., Pike BL., Vogel M., Amsler P., Rubin SM., van Leeuwen F., Guénolé A., van Attikum H., Thomä NH., Gasser SM. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation An N-terminal acidic region of Sgs1 interacts with Rpa70 and recruits Rad53 kinase to stalled forks. An N-terminal acidic region of Sgs1 interacts with Rpa70 and recruits Rad53 kinase to stalled forks. EMBO J. (0261-4189) 2012 imex curation Hegnauer AM., Hustedt N., Shimada K., Pike BL., Vogel M., Amsler P., Rubin SM., van Leeuwen F., Guénolé A., van Attikum H., Thomä NH., Gasser SM. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast anti tag coip anti tag coimmunoprecipitation anti tag coip experimental particp experimental participant identification experimental particp Hegnauer AM., Hustedt N., Shimada K., Pike BL., Vogel M., Amsler P., Rubin SM., van Leeuwen F., Guénolé A., van Attikum H., Thomä NH., Gasser SM. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation An N-terminal acidic region of Sgs1 interacts with Rpa70 and recruits Rad53 kinase to stalled forks. An N-terminal acidic region of Sgs1 interacts with Rpa70 and recruits Rad53 kinase to stalled forks. EMBO J. (0261-4189) 2012 imex curation Hegnauer AM., Hustedt N., Shimada K., Pike BL., Vogel M., Amsler P., Rubin SM., van Leeuwen F., Guénolé A., van Attikum H., Thomä NH., Gasser SM. Only protein-protein interactions yeast Saccharomyces cerevisiae Baker's yeast 2 hybrid two hybrid 2H 2h Gal4 transcription regeneration two-hybrid yeast two hybrid 2-hybrid classical two hybrid classical two hybrid 2H two-hybrid 2-hybrid Gal4 transcription regeneration 2h yeast two hybrid Y2H Y-2H 2 hybrid predetermined predetermined participant predetermined Hegnauer AM., Hustedt N., Shimada K., Pike BL., Vogel M., Amsler P., Rubin SM., van Leeuwen F., Guénolé A., van Attikum H., Thomä NH., Gasser SM. EMBO J. (0261-4189) 2012 Only protein-protein interactions imex curation rfa1_yeast Replication factor A protein 1 RFA1 BUF2 YAR007C FUN3 Replication protein A 69 kDa DNA-binding subunit Single-stranded DNA-binding protein DNA-binding protein BUF2 RPA1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MSSVQLSRGDFHSIFTNKQRYDNPTGGVYQVYNTRKSDGANSNRKNLIMISDGIYHMKALLRNQAASKFQSMELQRGDIIRVIIAEPAIVRERKKYVLLVDDFELVQSRADMVNQTSTFLDNYFSEHPNETLKDEDITDSGNVANQTNASNAGVPDMLHSNSNLNANERKFANENPNSQKTRPIFAIEQLSPYQNVWTIKARVSYKGEIKTWHNQRGDGKLFNVNFLDTSGEIRATAFNDFATKFNEILQEGKVYYVSKAKLQPAKPQFTNLTHPYELNLDRDTVIEECFDESNVPKTHFNFIKLDAIQNQEVNSNVDVLGIIQTINPHFELTSRAGKKFDRRDITIVDDSGFSISVGLWNQQALDFNLPEGSVAAIKGVRVTDFGGKSLSMGFSSTLIPNPEIPEAYALKGWYDSKGRNANFITLKQEPGMGGQSAASLTKFIAQRITIARAQAENLGRSEKGDFFSVKAAISFLKVDNFAYPACSNENCNKKVLEQPDGTWRCEKCDTNNARPNWRYILTISIIDETNQLWLTLFDDQAKQLLGVDANTLMSLKEEDPNEFTKITQSIQMNEYDFRIRAREDTYNDQSRIRYTVANLHSLNYRAEADYLADELSKALLA 7EB8DFA6910EF8A8 7EB8DFA6910EF8A8 sgs1_yeast ATP-dependent helicase SGS1 SGS1 TPS1 YMR190C YM9646.02C Helicase TPS1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MVTKPSHNLRREHKWLKETATLQEDKDFVFQAIQKHIANKRPKTNSPPTTPSKDECGPGTTNFITSIPASGPTNTATKQHEVMQTLSNDTEWLSYTATSNQYADVPMVDIPASTSVVSNPRTPNGSKTHNFNTFRPHMASSLVENDSSRNLGSRNNNKSVIDNSSIGKQLENDIKLEVIRLQGSLIMALKEQSKLLLQKCSIIESTSLSEDAKRLQLSRDIRPQLSNMSIRIDSLEKEIIKAKKDGMSKDQSKGRSQVSSQDDNIISSILPSPLEYNTSSRNSNLTSTTATTVTKALAITGAKQNITNNTGKNSNNDSNNDDLIQVLDDEDDIDCDPPVILKEGAPHSPAFPHLHMTSEEQDELTRRRNMRSREPVNYRIPDRDDPFDYVMGKSLRDDYPDVEREEDELTMEAEDDAHSSYMTTRDEEKEENELLNQSDFDFVVNDDLDPTQDTDYHDNMDVSANIQESSQEGDTRSTITLSQNKNVQVILSSPTAQSVPSNGQNQIGVEHIDLLEDDLEKDAILDDSMSFSFGRQHMPMSHSDLELIDSEKENEDFEEDNNNNGIEYLSDSDLERFDEERENRTQVADIQELDNDLKIITERKLTGDNEHPPPSWSPKIKREKSSVSQKDEEDDFDDDFSLSDIVSKSNLSSKTNGPTYPWSDEVLYRLHEVFKLPGFRPNQLEAVNATLQGKDVFVLMPTGGGKSLCYQLPAVVKSGKTHGTTIVISPLISLMQDQVEHLLNKNIKASMFSSRGTAEQRRQTFNLFINGLLDLVYISPEMISASEQCKRAISRLYADGKLARIVVDEAHCVSNWGHDFRPDYKELKFFKREYPDIPMIALTATASEQVRMDIIHNLELKEPVFLKQSFNRTNLYYEVNKKTKNTIFEICDAVKSRFKNQTGIIYCHSKKSCEQTSAQMQRNGIKCAYYHAGMEPDERLSVQKAWQADEIQVICATVAFGMGIDKPDVRFVYHFTVPRTLEGYYQETGRAGRDGNYSYCITYFSFRDIRTMQTMIQKDKNLDRENKEKHLNKLQQVMAYCDNVTDCRRKLVLSYFNEDFDSKLCHKNCDNCRNSANVINEERDVTEPAKKIVKLVESIQNERVTIIYCQDVFKGSRSSKIVQANHDTLEEHGIGKSMQKSEIERIFFHLITIRVLQEYSIMNNSGFASSYVKVGPNAKKLLTGKMEIKMQFTISAPNSRPSTSSSFQANEDNIPVIAQKSTTIGGNVAANPPRFISAKEHLRSYTYGGSTMGSSHPITLKNTSDLRSTQELNNLRMTYERLRELSLNLGNRMVPPVGNFMPDSILKKMAAILPMNDSAFATLGTVEDKYRRRFKYFKATIADLSKKRSSEDHEKYDTILNDEFVNRAAASSNGIAQSTGTKSKFFGANLNEAKENEQIINQIRQSQLPKNTTSSKSGTRSISKSSKKSANGRRGFRNYRGHYRGRK 0DC320B41756A3C3 0DC320B41756A3C3 rfa2_yeast Replication factor A protein 2 RFA2 BUF1 YNL312W N0368 Replication protein A 36 kDa subunit DNA-binding protein BUF1 protein protein yeast Saccharomyces cerevisiae Baker's yeast MATYQPYNEYSSVTGGGFENSESRPGSGESETNTRVNTLTPVTIKQILESKQDIQDGPFVSHNQELHHVCFVGVVRNITDHTANIFLTIEDGTGQIEVRKWSEDANDLAAGNDDSSGKGYGSQVAQQFEIGGYVKVFGALKEFGGKKNIQYAVIKPIDSFNEVLTHHLEVIKCHSIASGMMKQPLESASNNNGQSLFVKDDNDTSSGSSPLQRILEFCKKQCEGKDANSFAVPIPLISQSLNLDETTVRNCCTTLTDQGFIYPTFDDNNFFAL F6087501E4E28CC1 F6087501E4E28CC1 rad53_yeast Serine/threonine-protein kinase RAD53 RAD53 MEC2 SAD1 Serine-protein kinase 1 SPK1 P2588 YPL153C CHEK2 homolog protein protein yeast Saccharomyces cerevisiae Baker's yeast MENITQPTQQSTQATQRFLIEKFSQEQIGENIVCRVICTTGQIPIRDLSADISQVLKEKRSIKKVWTFGRNPACDYHLGNISRLSNKHFQILLGEDGNLLLNDISTNGTWLNGQKVEKNSNQLLSQGDEITVGVGVESDILSLVIFINDKFKQCLEQNKVDRIRSNLKNTSKIASPGLTSSTASSMVANKTGIFKDFSIIDEVVGQGAFATVKKAIERTTGKTFAVKIISKRKVIGNMDGVTRELEVLQKLNHPRIVRLKGFYEDTESYYMVMEFVSGGDLMDFVAAHGAVGEDAGREISRQILTAIKYIHSMGISHRDLKPDNILIEQDDPVLVKITDFGLAKVQGNGSFMKTFCGTLAYVAPEVIRGKDTSVSPDEYEERNEYSSLVDMWSMGCLVYVILTGHLPFSGSTQDQLYKQIGRGSYHEGPLKDFRISEEARDFIDSLLQVDPNNRSTAAKALNHPWIKMSPLGSQSYGDFSQISLSQSLSQQKLLENMDDAQYEFVKAQRKLQMEQQLQEQDQEDQDGKIQGFKIPAHAPIRYTQPKSIEAETREQKLLHSNNTENVKSSKKKGNGRFLTLKPLPDSIIQESLEIQQGVNPFFIGRSEDCNCKIEDNRLSRVHCFIFKKRHAVGKSMYESPAQGLDDIWYCHTGTNVSYLNNNRMIQGTKFLLQDGDEIKIIWDKNNKFVIGFKVEINDTTGLFNEGLGMLQEQRVVLKQTAEEKDLVKKLTQMMAAQRANQPSASSSSMSAKKPPVSDTNNNGNNSVLNDLVESPINANTGNILKRIHSVSLSQSQIDPSKKVKRAKLDQTSKGPENLQFS 84A9612229CA72D1 84A9612229CA72D1 rfa1-sgs1 3 Rpa70 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Sgs1 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association homomint domino mint Similarly, we mapped the interaction site on Rpa70 by fusing Rpa70 subdomains to the LexA-DBD and monitoring their interaction with the Sgs1(290 1180)-B42-AD fusion. The highest b-galactosidase activity was measured for the N-terminal oligonucleotide binding fold (N-OB) of Rpa70 without the linker region (Supplementary Figure S1A). f1c f1d sf1a rfa2-sgs1 3 Rpa32 6 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Sgs1 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain physical association physical association sf1b homomint mint sgs1-rad53 3 RAD53 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Sgs1 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell fusion protein fusion protein fusion protein undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined binding site binding region binding-associated region binding region deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain physical association physical association f6a homomint domino mint We detect a strong interaction between the core Sgs1 fragment (290 1180) and the FHA1 domain of Rad53, which is entirely lost upon deletion of aa 404 604 (sgs1-r1; Figure 6A). The Sgs1 subdomain aa 292 661 was sufficient to mediate binding to the Rad53 FHA1 domain, and again, deletion of aa 404 604 ablated interaction with Rad53 FHA1 (Figure 6A). rfa1-sgs1-1 1 Rpa70 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Sgs1 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 30 32 direct interaction direct interaction f1e homomint domino mint rfa1-sgs1-2 1 Rpa70 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain Sgs1 5 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 35 37 direct interaction direct interaction f1f homomint domino mint rad53-sgs1 1 Sgs1 5 peptide synthesis peptide synthesis experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified phosphorylated phosres phosphorylated residue PhosRes Phospho Phosphorylation Phosphorylation (O of Serine, Threonine, Tyrosine and Aspartate, N epsilon of Lysine) certain certain sequence position certain certain certain sequence position certain prerequisite-ptm binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain RAD53 7 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role neutral component neutral component purified purified binding site binding region binding-associated region binding region certain certain sequence position certain certain certain sequence position certain 43 41 direct interaction direct interaction f6b homomint domino mint sgs1-rfa1 2 Sgs1 5 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:19:34 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation decreasing' 404-604 myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Rpa70 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f2c homomint mint rfa1-sgs1-3 2 Rpa70 4 predetermined predetermined participant predetermined experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell ha tag ha tag ha tag YPYDVPDYA epitope tag YPYDVPDYA epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined Sgs1 5 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined undefined mutation mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing deletion analysis deletion analysis certain certain sequence position certain certain certain sequence position certain Wed Jul 06 16:21:12 BST 2016 Sequence change details about this feature cannot be ascertained or do not fit with the current version of the referenced protein, so they have been deleted as a result of our quality control procedures. The original label was 'mutation decreasing' 404-604 physical association physical association sf2 homomint mint sgs1-rad53-1 2 Sgs1 5 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role bait bait over-expressed over expressed level over-expressed living cell living cell P35187:p.Thr585Ala mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T A Tue Jul 12 10:17:43 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 19:22:38 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr585ala' Tue Jun 30 17:46:32 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr585Ala' P35187:p.Ser470Ala mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Tue Jul 12 10:17:46 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 19:34:42 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser470ala' Tue Jun 30 17:32:06 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser470Ala' P35187:p.Thr451Ala mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain T A Tue Jul 12 10:55:29 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 15:52:21 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'thr451ala' Tue Jun 30 16:45:12 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Thr451Ala' P35187:p.Ser482Ala mutation decreasing interaction mutation decreasing mutation decreasing interaction hotspot hotspot mutation decreasing mutation analysis mutation analysis certain certain sequence position certain certain certain sequence position certain S A Tue Jul 12 10:55:29 BST 2016 This feature has been corrected as a result of our quality control procedures. The original label was 'mutation decreasing' Wed May 02 16:19:55 BST 2018 This feature has been corrected as a result of our quality control procedures. The original label was 'ser482ala' Tue Jun 30 20:13:23 BST 2020 This feature has been corrected as a result of our quality control procedures. The original label was 'p.Ser482Ala' myc tag myc tag myc tag EUKLISEED epitope tag EUKLISEED epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined RAD53 7 anti tag western anti tag western blot anti tag western experimental particp experimental participant identification experimental particp unspecified role unspecified role prey prey over-expressed over expressed level over-expressed living cell living cell v5 tag v5 tag v5 tag GKPIPNPLLGLDST epitope tag GKPIPNPLLGLDST epitope tag undetermined undetermined sequence position undetermined undetermined undetermined sequence position undetermined physical association physical association f6c homomint domino mint